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Swine production workers are exposed simultaneously to multiple contaminants. Occupational exposure to aflatoxin B1 (AFB1) in Portuguese swine production farms has already been reported. However, besides AFB1, data regarding fungal contamination showed that exposure to other mycotoxins could be expected in this setting. The present study aimed to characterize the occupational exposure to multiple mycotoxins of swine production workers. To provide a broad view on the burden of contamination by mycotoxins and the workers&rsquo, exposure, biological (urine) samples from workers (n = 25) and 38 environmental samples (air samples, n = 23, litter samples, n = 5, feed samples, n = 10) were collected. The mycotoxins biomarkers detected in the urine samples of the workers group were the deoxynivalenol-glucuronic acid conjugate (60%), aflatoxin M1 (16%), enniatin B (4%), citrinin (8%), dihydrocitrinone (12%) and ochratoxin A (80%). Results of the control group followed the same pattern, but in general with a lower number of quantifiable results (&lt, LOQ). Besides air samples, all the other environmental samples collected presented high and diverse contamination, and deoxynivalenol (DON), like in the biomonitoring results, was the most prominent mycotoxin. The results demonstrate that the occupational environment is adding and contributing to the workers&rsquo, total exposure to mycotoxins, particularly in the case of DON. This was confirmed by the biomonitoring data and the high contamination found in feed and litter samples. Furthermore, he followed multi-biomarker approach allowed to conclude that workers and general population are exposed to several mycotoxins simultaneously. Moreover, occupational exposure is probably described as being intermittent and with very high concentrations for short durations. This should be reflected in the risk assessment process.

Mycotoxins are toxic mold metabolites that can persist in environment long after the fungi species responsible for their production disappear. Critical workplace for mycotoxins presence has already been studied and nowadays it is possible to recognize that exposure to mycotoxins through inhalation occurs due to their presence in dust. This study aimed to assess occupational co-exposure to multiple mycotoxins in a fresh bread dough company, an occupational setting not studied until now. Occupational exposure assessment to mycotoxins was done using a LC-MS/MS urinary multi-biomarker approach. Twenty-one workers and nineteen individuals that were used as controls participated in the study. Workers/controls (spot-urine) and environment (settled dust) samples were collected and analyzed. Concerning workers group, DON-GlcA, and OTA were the most prevalent biomarkers (&gt, LOD), 66% and 90.5%, respectively. In the control group, OTA was also one of the most detected (68%) followed by CIT (58%) and DON-GlcA (58%). DON was the mycotoxin measured in high amounts in the settled dust sample (58.2 ng/g). Both workers and controls are exposed to several mycotoxins simultaneously. The workers group, due to their high contact with flour dust, revealed a higher exposure to DON. Considering these results, risk management measures must be applied including specific and adequate health surveillance programs in order to avoid exposure and consequently the associated health consequences.

Scope In this study, human exposure to the mycotoxin ochratoxin A (OTA) and its thermal degradation product 2’R-ochratoxin A (2’R-OTA, previously named as 14R-Ochratoxin A [22]) through coffee consumption was assessed. LC-MS/MS and the dried blood spot (DBS) technique were used for the analysis of blood samples from coffee and noncoffee drinkers (n = 50), and food frequency questionnaires were used to document coffee consumption. Methods and results For the detection of OTA and 2’R-OTA in blood, a new sensitive and efficient sample preparation method based on DBS was established and validated. Using this technique 2’R-OTA was for the first time detected in biological samples. Comparison between coffee drinkers and noncoffee drinkers showed for the first time that 2’R-OTA was only present in blood from the first group while OTA could be found in both groups in a mean concentration of 0.21 μg/L. 2’R-OTA mean concentration was 0.11 μg/L with a maximum concentration of 0.414 μg/L. Thus, in average 2’R-OTA was approx. half the concentration of OTA but in some cases even exceeded OTA levels. No correlation between the amounts of coffee consumption and OTA or 2’R-OTA levels was observed. Conclusion The results of this study revealed for the first time a high exposure of coffee consumers to 2’R-OTA, a compound formed from OTA during coffee roasting. Since little information is available regarding toxicity and possible carcinogenicity of this compound, further OTA monitoring in blood including 2’R-OTA is advisable.

The waste management occupational environment is recognized by the simultaneous presence of several substances and biologic agents. Therefore, workers are exposed simultaneously to multiple contaminants. Occupational exposure to aflatoxin B1 in one Portuguese waste sorting plant was already reported. However, besides this mycotoxin, data regarding fungal contamination showed that exposure to other mycotoxins could be expected. A study was developed to analyze if exposure to other mycotoxins besides aflatoxin B1 was occurring in the workers from the waste sorting plant previously assessed and to discuss how these findings need to be considered in the risk assessment process. In addition to aflatoxin B1 detected previously by ELISA, two additional mycotoxins and one mycotoxin degradation product were detected and quantified by a multi-mycotoxin HPLC-MS/MS approach: Enniatin B and ochratoxin A as well as 2’R-ochratoxin A. Besides the confirmation of co-exposure to several mycotoxins, results probably indicate different exposure routes for the mycotoxins reported.

In this study, a rapid multi-mycotoxin approach was developed for biomonitoring and quantification of 27 important mycotoxins and mycotoxin metabolites in human blood samples. HPLC-MS/MS detection was used for the analysis of dried serum spots (DSS) and dried blood spots (DBS). Detection of aflatoxins (AFB1, AFB2, AFG1, AFG2, AFM1), trichothecenes (deoxynivalenol, DON; DON-3-glucoronic acid, DON-3-GlcA; T-2; HT-2; and HT-2-4-GlcA), fumonisin B1 (FB1), ochratoxins (OTA and its thermal degradation product 2’R-OTA; OTα; 10-hydroxychratoxin A, 10-OH-OTA), citrinin (CIT and its urinary metabolite dihydrocitrinone, DH-CIT), zearalenone and zearalanone (ZEN, ZAN), altenuene (ALT), alternariols (AOH; alternariol monomethyl ether, AME), enniatins (EnA, EnA1, EnB, EnB1) and beauvericin (Bea) was validated for two matrices, serum (DSS), and whole blood (DBS). HPLC-MS/MS analysis showed signal suppression as well as signal enhancement due to matrix effects. However, for most analytes LOQs in the lower pg/mL range and excellent recovery rate were achieved using matrix-matched calibration. Besides validation of the method, the analyte stability in DBS and DSS was also investigated. Stability is a main issue for some analytes when the dried samples are stored under common conditions at room temperature. Nevertheless, the developed method was applied to DBS samples of a German cohort (n = 50). Besides positive findings of OTA and 2’R-OTA, all samples were positive for EnB. This methodical study establishes a validated multi-mycotoxin approach for the detection of 27 mycotoxins and metabolites in dried blood/serum spots based on a fast sample preparation followed by sensitive HPLC-MS/MS analysis. Graphical Abstractᅟ Electronic supplementary material The online version of this article (doi:10.1007/s00216-017-0279-9) contains supplementary material, which is available to authorized users.